Evidence of chemical bonding at biomaterial-hard tissue interfaces

For many years, glass-polyalkenoate cements have been described as possessing the unique properties of self-adherence to human hard tissues, such as bones or teeth. However, direct experimental evidence to prove the existence of chemical bonding has not been advanced. X-ray Photoelectron Spectroscopy (XPS) was used to analyze the chemical interaction of a synthesized polyalkenoic acid with enamel and synthetic hydroxyapatite. For both enamel and hydroxyapatite, the peak representing the carboxyl groups of the polyalkenoic acid was detected to have significantly shifted to a lower binding energy. De-convolution of this shifted peak disclosed two components with a peak representing unreacted carboxyl groups and a peak suggesting chemical bonding to hydroxyapatite. On average, 67.5% of the carboxyl groups of the polyalkenoic acid were measured to have bonded to hydroxyapatite. XPS of hydroxyapatite also disclosed its surface to be enriched in calcium and decreased in phosphorus, indicating that phosphorus was extracted at a relatively higher rate than calcium. Analysis of these data supports the mechanism in which carboxylic groups replace phosphate ions (PO4(3-)) of the substrate and make ionic bonds with calcium ions of hydroxyapatite. It is concluded that an ultrathin layer of a polyalkenoic acid can be prepared on a hydroxyapatite-based substrate by careful removal of non-bonded molecules. With this specimen-processing method, XPS not only provided direct evidence of chemical bonding, but also enabled us to quantify the percentages of functional groups of the polyalkenoic acids that bonded to calcium of hydroxyapatite.     

In situ tracing the process of human enamel demineralization by electrochemical impedance spectroscopy (EIS)

OBJECTIVE: The purpose of this study was to in situ characterize the demineralization (namely dissolution of hydroxyapatite) on the surface of the human enamel using electrochemical impedance spectroscopy (EIS) technology. METHODS: Fresh human third molars extracted without visible evidence of caries, were used in this study. After they were immersed in a demineralizing solution prepared from lactic acid and carboxy methyl cellulose sodium (Na-CMC) buffering at pH 4, demineralization happened on their surfaces. EIS of the specimens were measured at a series of immersed interval. X-ray diffractometer (XRD) were used to distinguish the microstructure of the surface layer of the specimens. The depositions that appeared in the demineralizing solution after 46 h immersion were analyzed by fourier transform infrared spectrometer (FTIR). RESULTS: XRD analysis revealed that the percentage of intensity (I%) of HAP gradually decreased with the elapsed immersing time, which indicated the dissolution of HAP columns of enamel. Nyquist spectra were fitted with an equivalent circuit characterized by some parameters, such as Q and R(p) (error<0.1%). Changes of the parameters' values revealed that the rate of demineralization accelerated at the forepart of the demineralization, but slowed down beyond 70 h immersion. CONCLUSIONS: The results suggested that EIS was proved to be a useful method for in situ investigating and in vivo detecting the demineralization of the enamel.     

Combinations of etchants, composite resins, and bracket systems: an important choice in orthodontic bonding procedures.

The objectives of this investigation were: (1) to compare the shear bond strengths (SBS) of metal, ceramic, and plastic brackets using different concentrations of maleic and phosphoric acid gels and aqueous solutions, and (2) to determine if a relationship exists between the type of acid etchant and the location of resin after debonding. A sample of 210 bovine incisors was divided among three different bracket groups (Victory series metal, Transcend 6000 ceramic, Spirit MB plastic). Prior to bonding, enamel was acid-etched using 37% phosphoric acid (H3PO4) gel and aqueous solution, 10% maleic acid gel and aqueous solution, 10% H3PO4 gel and aqueous solution, or 2% H3PO4 aqueous solution. SBS testing and the adhesive remnant index (ARI) score provided insight into the effects of the bonding process on enamel. Resin tags associated with each etchant type were inspected under scanning electron microscopy (SEM). Statistical analyses (level of significance, p = 0.05) of the data showed significant differences among groups. It was concluded that specific acid-composite-bracket combinations are recommended for use in clinical orthodontic practice in order to achieve efficient bonding.     

Crystalline structure of dental enamel after Ho:YLF laser irradiation

Irradiation of teeth with lasers using specific wavelengths and energy densities produces surface melting. This effect has been already applied to different procedures such as caries prevention and hypersensitivity reduction. The aim of this study is to characterize the crystalline structure of bovine enamel after holmium laser irradiation. A holmium laser (Ho:YLF) with emission wavelength of 2065 nm was used. Enamel tissues were irradiated in ablative regime and their structures before and after irradiation were analyzed using the powder X-ray diffraction technique. The X-ray diffraction patterns of non-irradiated enamel correspond to carbonated hydroxyapatite and those produced by irradiated samples indicate the existence of a mixture of two crystalline phases: hydroxyapatite and tetracalcium phosphate. The structural characteristics of holmium irradiated enamel were compared with those of the same tissue irradiated with other lasers.     

Synchrotron radiation microbeam X-ray fluorescence analysis of zinc concentration in remineralized enamel in situ

Objective

Remineralization is an indispensable phenomenon during the natural healing process of enamel decay. The incorporation of zinc (Zn) into enamel crystal could accelerate this remineralization. The present study was designed to investigate the concentration and distribution of Zn in remineralized enamel after gum chewing.

Methods

The experiment was performed at the Photon Factory. Synchrotron radiation was monochromatized and X-rays were focused into a small beam spot. The X-ray fluorescence (XRF) from the sample was detected with a silicon (Si) (lithium (Li)) detector. X-ray beam energy was tuned to detect Zn. The examined samples were small enamel fragments remineralized after chewing calcium phosphate-containing gum in situ. The incorporation of Zn atom into hydroxyapatite (OHAP), the main component of enamel, was measured using Zn K-edge extended X-ray absorption fine structure (EXAFS) with fluorescence mode at the SPring-8.

Results

A high concentration of Zn was detected in a superficial area 10-μm deep of the sectioned enamel after gum chewing. This concentration increased over that in the intact enamel. The atomic distance between Zn and O in the enamel was calculated using the EXAFS data. The analyzed atomic distances between Zn and O in two sections were 0.237 and 0.240 nm.

Conclusion

The present experiments suggest that Zn is effectively incorporated into remineralized enamel through the physiological processes of mineral deposition in the oral cavity through gum-chewing and that Zn substitution probably occurred at the calcium position in enamel hydroxyapatite.     

The formation and remineralization of artificial white spot lesions: a constant composition approach

Artificial white spot lesions have been prepared on bovine enamel surfaces by the controlled addition of lactate buffer containing methylhydroxydiphosphonate ions. The rates of remineralization have been measured using a constant composition method in solutions of calcium phosphate of low supersaturation. Deposition of new hydroxyapatite solid phase takes place exclusively within the white spot lesion at a rate appreciably slower than that of an acid-etched enamel surface, probably due to the presence of methylhydroxydiphosphonate located within the intact surface region of the lesion.     

Enamel dissolution in citric acid as a function of calcium and phosphate concentrations and degree of saturation with respect to hydroxyapatite

The aim of this study was to investigate enamel dissolution in citric acid solutions as a function of solution calcium and phosphate concentrations and degree of saturation with respect to hydroxyapatite (DSHA). The primary relevance of the study is the development of soft drinks with reduced erosive potential. Nanoindentation was used to investigate changes in the hardness of polished human enamel surfaces after 120 s and 300 s exposure to solutions with pH 3.30 and a range of calcium and phosphate concentrations. All solutions were undersaturated with respect to hydroxyapatite, with 0.000 < or = DSHA < or = 0.295. A complex dependence of enamel softening on calcium concentration was observed. Substantial enamel softening occurred in solutions with calcium concentrations equal to or less than 120 mm (DSHA approximately 0.104), but there was little or no statistically significant softening of the enamel for calcium concentrations over 120 mm. This condition may be applicable to soft drink formulation. Furthermore, solutions with DSHA = 0.101 and different calcium/phosphate ratios resulted in different degrees of softening of the enamel. Hence, contrary to assumptions made in many models, enamel dissolution is not simply a function of DSHA, and individual calcium and phosphate concentrations are critical.     

The demonstration of a dicalcium phosphate stabilizing factor in human saliva

Dicalcium phosphate dihydrate (DCPD) was equilibrated in H₂O, buffer solutions at pH 7.4, saliva samples and their ultrafiltrates and suspensions of cellular residue obtained from centrifiugation of whole saliva. The supernatants were analysed chemically for calcium phosphate and pH after 48 hr and the solids examined by X-ray diffraction. DCPD suspended in saliva remained unchanged in 18 out of 21 duct salivas and in 12 out of 26 samples of whole saliva, while the more basic calcium phosphate, octacalcium phosphate, was formed in 43 out of 47 ultrafiltrates and in water and buffer solutions. DCPD also remained unchanged in suspensions of salivary residue. If saliva was pretreated with ground enamel powder or synthetic hydroxyapatite prior to DCPD equilibrium, more basic calcium phosphate would form. It is concluded that saliva contains a DCPD stabilizing factor which is non-ultrafiltrable and which may be adsorbed on to enamel powder, synthetic hydroxyapatite and salivary cellular residue.     

釉质仿生矿化模型的研究

目的：构建釉质生物矿化的模型,包括有机基质模板（类釉原蛋白寡肽序列）的建立和无机离子供体（包裹钙磷离子的温度敏感性脂质体）的合成,体外实现类釉质样结构的再矿化。方法：首先通过标准固相法合成所需“类釉原蛋白”寡肽[（Gln?Pro?Ala）4?Thr?Lys?Arg?Glu?Glu?Val?Asp],并用CaCl2溶液诱导其进行自组装;其次采用二棕榈酰磷脂酰胆碱和二肉豆蔻卵磷脂为原料,通过相交融合法分别合成包裹钙、磷离子的温度敏感性脂质体;最后在37℃时,将酸蚀后的牙片浸泡在包裹钙、磷离子的脂质体与寡肽的混悬液中,作为实验组。将酸蚀后的牙片浸泡在包裹钙磷离子的脂质体混悬液中,作为对照组,促进脱矿后的釉质再矿化。矿化后的牙片通过扫描电镜（ SEM）、傅立叶变换红外光谱仪（ FTIR）和X射线衍射仪（ XRD）进行表征。结果：实验组的脱矿釉质表面均匀有序的沉积了一层釉质样的羟基磷灰石晶体（ HA）结构,而对照组只沉积了较少量无序的HA晶体。结论：通过类釉原蛋白寡肽有机矿化模板的建立,以及仿生釉质矿化过程中钙、磷离子的输送,构建了釉质仿生矿化模型,并实现了脱矿釉质表面类釉质样微结构的再生。     

Analysis of chemical interaction of 4-MET with hydroxyapatite using XPS

Each dental adhesive contains a specific functional monomer that determines its actual adhesive performance to tooth tissue. 4-methacryloxyethyl trimellitic acid (4-MET) is well-known as one of the functional monomers mostly available and consequently widely used in commercial adhesives. We therefore characterized the chemical interaction of 4-MET with hydroxyapatite (HAp) using X-ray Photoelectron Spectroscopy (XPS). XPS revealed that the peak representing -COO- of 4-MET shifted to a lower binding energy, when 4-MET was adsorbed onto HAp. Deconvolution of this shifted peak disclosed two components with a peak representing unreacted carboxyl groups and ester groups, and a peak suggesting chemical bonding of other carboxyl groups to Ca of HAp. XPS spectra of HAp treated with 4-MET also disclosed the surface to be enriched in calcium and decreased in phosphorus, indicating that phosphorus was extracted at a relatively higher rate than calcium. It can thus be concluded that true chemical bonding of 4-MET with calcium present in HAp occurred, as it was proven using XPS.     

Effect of soluble calcium on fluoride uptake by enamel from sodium monofluorophosphate

The fluoride uptake by sound, acid-etched and artificial white spot lesions treated with sodium monofluorophosphate in dentifrice slurries, abrasive slurries, or calcium chloride solutions was determined. Fluoride uptake by sound enamel decreased in the presence of soluble calcium; however, uptake by etched and white spot enamel was greatly enhanced. The optimum Ca:MFP molar ratio was 0.5. Because fluorine-containing dentifrices probably inhibit dental caries by interaction with white spots or at the initial stage of acid-attack, the soluble calcium contained in saliva or a dentifrice might enhance the anti-caries effect of monofluorophosphate.     

Localized fluoride release from fluorine-carrying polyphosphonates.

Fluoride release from fluorine-carrying copolymers of vinylphosphonate induced by calcium apatite and tooth enamel has been investigated. Fluoride ions were determined potentiometrically in the study of calcium hydroxyapatite, and Auger spectroscopy was used to study the fluoride release to enamel. The adsorbed copolymer of vinylphosphonic acid and vinylphosphonyl thiofluoride was shown to release fluoride to calcium hydroxyapatite and to enamel. The oxygen analog has shown a similar behavior with calcium hydroxyapatite, but not with tooth enamel. The results suggest a potential application of such copolymers as caries preventive agents which combine the effects of polyphosphonates and of fluoride ions.     

Bond strength of resin to acid-etched dentin studied by 13C NMR: interaction between N-methacryloyl-omega-amino acid primer and dentinal collagen

The application of the hydrophilic methacrylate primer to acid-etched dentin increases the bond strength of the resin to the acid-etched dentin. However, the mechanism for the bonding of the resin to dentinal collagen through the primer remains to be determined. Before a more effective adhesive primer can be developed, we must understand the adsorption behavior of the primer to the dentinal collagen. The purpose of this study was to determine how 5 derivatives of N-methacryloyl-omega-amino acid (NMomegaA) primers enhance the bonding of the resin to acid-etched dentin. The interaction between the NMomegaA primers and dentinal collagen was studied by the 13C NMR technique, including the observation of spin-lattice relaxation times, T1. When the dentinal collagen was dispersed into the NMomegaA solution, the T1 values of the two carbonyl carbons attributed to the amide and the carboxylic acid in the NMomegaA molecule decreased dramatically. This result was due to the interaction between the amide group and the carboxylic acid group in the NMomegaA molecule and the dentinal collagen molecule. The T1 values of these carbonyl carbons decreased when the number of methylene groups in the NMomegaA molecule increased. The interaction became stronger as the number of methylene groups in the NMomegaA molecule was increased. Further, the bond strength of the resin to the acid-etched dentin primed with NMomegaA increased with a decrease in the T1 value of the amide carbonyl carbon. The strength of the interaction of the NMomegaA primer to the dentinal collagen molecule, determined by the 13C NMR technique, showed a direct correlation with the bond strength of the resin to acid-etched dentin that was treated with the NMomegaA primer.     

Acid resistance of erbium-doped yttrium aluminum garnet laser-treated and phosphoric acid-etched enamels

OBJECTIVE: To compare the effects of erbium-doped yttrium aluminum garnet (Er:YAG) laser ablation and of phosphoric acid etching on the in vitro acid resistance of bovine enamel. MATERIALS AND METHODS: Teeth were polished to make the surface flat. The polished enamel was either etched with 37% phosphoric acid for 30 seconds or ablated with a single 33 J/cm2 pulse from an Er:YAG laser. The control specimens were free from acid etching and laser ablation. Changes in crystal structure, dissolved mineral (calcium [Ca] and phosphorus [P]) contents, and calcium distribution in the enamel subsurface after a pH-cycling process were evaluated. RESULTS: After laser treatment, poor crystal structures improved without forming any new phases, such as tricalcium phosphates. Among the tested enamels, dissolved mineral contents were significantly different (P < .05). Er:YAG laser-treated enamels had the lowest mineral dissolution (Ca, 13.78 ppm; P, 6.33 ppm), whereas phosphoric acid-etched enamels had the highest (Ca, 15.90 ppm; P, 7.33 ppm). The reduction rate and reduced depth of calcium content along the subsurface were lowest in Er:YAG laser-treated enamels. CONCLUSION: The Er:YAG laser-treated enamels are more acid resistant to acid attack than phosphoric acid-etched enamels.     

8DSS peptide induced effective dentinal tubule occlusion in vitro

Objective

Eight repetitive nucleotide sequences of aspartate–serine–serine (8DSS) derived from dentin phosphoprotein (DPP) has been proved to be a good remineralization agency. In this study, 8DSS peptide was employed to induce dentinal tubule occlusion.

Characterization of enamel with variable caries risk

Small angle X-ray scattering (SAXS), X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR) have been used to look at enamel from nine premolars, three each from individuals in low, medium and high risk caries groups. Only SAXS was able to detect consistent differences between any of the groups. In enamel from the high caries risk group, the micropores between the hydroxyapatite crystals were laminar. In enamel from the low caries risk group, the micropores were cylindrical. Other parameters varied between teeth but were not correlated with caries risk.     

Laser-Raman spectroscopic study of the adhesive interface; analysis between 4-META/MMA-TBB resin and bovine or human dentin.

A study of the adhesive interface between 4-MET/MMA-TBB resin and hydroxyapatite or bovine enamel was reported. The present report is a continuation of that study. The possible chemical interaction between 4-methacryloxyethyl trimellitic acid (4-MET) and bovine or human dentin was examined by laser Raman spectroscopy. A 4-MET monomer solution was prepared by evaporating two thirds of the methyl methacrylate (MMA) in a commercial dentin adhesive. The solution was then applied to a dentin surface after treating the surface with an aqueous solution of 10% citric acid containing 3% ferric chloride. A salt formed on both bovine and human dentin surfaces. This salt was formed by the process we previously reported in which 4-MET formed a salt on the hydroxyapatite and bovine enamel. No evidence was observed of chemical reaction between 4-MET and any organic component in the dentin.     

钛合金种植体（Ti6Al4V）表面接枝丙烯酸-衣康酸共聚物增强生物矿化的研究

目的通过在钛种植体（Ti6Al4V）表面接枝丙烯酸-农康酸共聚物以促进钛合金种植体表面羟基磷灰石的沉积。方法用异丙醇钛（TIP0）作偶联剂,在经氢氧化钠处理的钛种植体表面接枝丙烯酸-衣康酸共聚物,然后,将其浸入SBF溶液中一周进行生物矿化。运用扫描电子显做镜、X-射线衍射仪、X-射线能谱仪对表面矿化物的形貌、结构和组成进行表征和分析。结果接枝丙烯酸-衣康酸共聚物的钛种植体表面经SBF溶液浸泡后,其表面形成连续的针状矿化物,X-射线衍射仪、X-射线能谱仪证实其结构为羟基磷灰石。结论钛表面接枝丙烯酸-衣康酸共聚物后,能加快羟基磷灰石在表面的沉积,并调整其晶体形貌。     

Adhesive bonding of various materials to hard tooth tissues. XIV. Enamel mordant selection assisted by ESCA (XPS).

Surface analysis by XPS (X-ray photoelection spectroscopy), also called ESCA (electron spectroscopy for chemical analysis), indicates that only certain cations are appreciably sorbed by enamel from an acid etching solution containing phosphoric acid and equimolar concentrations of candidate mordant salts.     

Dentin bonding--state of the art 1999

The adhesion of restorative materials to the hard components of tooth structure has been a goal pursued by many researchers ever since Buonocore established the foundation for adhesive and preventive dentistry. Based on the industrial use of phosphoric acid to obtain better adhesion of paints and resin coatings to metal surfaces, Buonocore proposed that phosphoric acid could be used to transform the surface of enamel to "render it more receptive to adhesion." Subsequent research indicated that the formation of taglike resin prolongations into the enamel microporosities was the leading bonding mechanism of resin to phosphoric acid-etched enamel. The enamel bonding agents of the 1960s and 1970s progressively evolved into complex multibottle or universal adhesives in the early 1990s, which were designed to bond to enamel, dentin, composite, amalgam, porcelain, and non-precious metal. Although bonding to enamel has been a dependable technique, bonding to dentin still represents an overwhelming task because dentin is a naturally wet organic tissue penetrated by a tubular maze containing the odontoblastic process, which communicates with the pulp. This intrinsic moisture may actually benefit the chemistry of the newest adhesive systems, which must be applied on moist dentin to be effective. In fact, the collapse of the collagen that occurs on air-drying may prevent the adhesive monomers from penetrating the network of nano-channels formed by the dissolution of hydroxyapatite crystals between collagen fibrils. In view of the complexity of the mechanisms associated with these mechanisms, the objective of this review article is to summarize the most recent concepts in dentin bonding.